Proliferation and differentiation of cells of multicellular organisms are controlled by hormones and polypeptide growth factors. These diffusable molecules allow cells to communicate with each other and act in concert to regulate cells and form organs, and to repair and regenerate damaged tissue. Examples of hormones and growth factors include the steroid hormones (e.g. estrogen, testosterone), parathyroid hormone, follicle stimulating hormone, the interleukins, platelet derived growth factor (PDGF), epidermal growth factor (EGF), granulocyte-macrophage colony stimulating factor (GM-CSF), erythropoietin (EPO) and calcitonin.
Hormones and growth factors influence cellular metabolism by binding to proteins. These proteins may be integral membrane proteins that are linked to signaling pathways within the cell, such as second messenger systems. Other classes of proteins that hormones and growth factors influence are soluble molecules, such as the transcription factors.
There is a continuing need to discover new hormones, growth factors and the like. The in vivo activities of these cytokines illustrates the enormous clinical potential of, and need for, other cytokines, cytokine agonists, and cytokine antagonists. The present invention addresses this need by providing such polypeptides for these and other uses that should be apparent to those skilled in the art from the teachings herein.
The present invention addresses this need by providing a novel polypeptide and related compositions and methods.
Within one aspect, the present invention provides an isolated polynucleotide that encodes a polypeptide comprising a sequence of amino acid residues that is at least 90% identical to an amino acid sequence selected from the group consisting of: (a) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 26 (Tyr), to amino acid number 235 (Ser); and (b) the amino acid sequence as shown in SEQ ID NO:2 from amino acid number 1 (Met), to amino acid number 235 (Ser). Within one embodiment, the isolated polynucleotide molecule is selected from the group consisting of: (a) polynucleotide molecules comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 194 to nucleotide 823; (b) polynucleotide molecules comprising a nucleotide sequence as shown in SEQ ID NO:1 from nucleotide 119 to nucleotide 823; and (c) polynucleotide molecules complementary to (a) or (b). Within another embodiment, the isolated polynucleotide disclosed above comprises nucleotide 1 to nucleotide 705 of SEQ ID NO:8. Within another embodiment, the isolated polynucleotide disclosed above consists of a sequence of amino acid residues that is at least 90% identical to an amino acid sequence as shown in SEQ ID NO:2 from amino acid number 26 (Tyr), to amino acid number 235 (Ser). Within another embodiment, the isolated polynucleotide disclosed above consists of a sequence of amino acid residues as shown in SEQ ID NO:2 from amino acid number 26 (Tyr), to amino acid number 235 (Ser) Within another embodiment, the isolated polynucleotide disclosed above encodes a polypeptide that contains motifs 1, 2, 3, 4 and 5 spaced apart from N-terminus to C-terminus in a configuration M1-{25-26}-M2-{15}-M3-{11}-M4-{34-36}-M5.
Within a second aspect, the present invention provides an expression vector comprising the following operably linked elements: a transcription promoter; a DNA segment encoding a z219a polypeptide that is at least 90% identical to an amino acid sequence as shown in SEQ ID NO:2 from amino acid number 26 (Tyr), to amino acid number 235 (Ser); and a transcription terminator, wherein the promoter is operably linked to the DNA segment, and the DNA segment is operably linked to the transcription terminator. Within one embodiment, the expression vector disclosed above, further comprises a secretory signal sequence operably linked to the DNA segment.
Within a third aspect, the present invention provides, a cultured cell into which has been introduced an expression vector as disclosed above, wherein the cell expresses the polypeptide encoded by the DNA segment.
Within a fourth aspect, the present invention provides a DNA construct encoding a fusion protein, the DNA construct comprising: a first DNA segment encoding a polypeptide that is at least 90% identical to a sequence of amino acid residues 1 (Met) through 25 (Gly) of SEQ ID NO:2; and second DNA segment encoding an additional polypeptide, wherein the first and second DNA segments are connected in-frame; and encode the fusion protein.
Within another aspect, the present invention provides an isolated polypeptide comprising a sequence of amino acid residues that is at least 90% identical to an amino acid sequence selected from the group consisting of: (a) polypeptide molecules comprising an amino acid sequence as shown in SEQ ID NO:2 from amino acid number 26 (Tyr) to amino acid number 235 (Ser) of SEQ ID NO:2; (b) polypeptide molecules comprising an amino acid sequence as shown in SEQ ID NO:2 from amino acid residue number 1 (Met) to amino acid residue number 235 (Ser). Within one embodiment, the isolated polypeptide disclosed above consists of a sequence of amino acid residues that is at least 90% identical to an amino acid sequence as shown in SEQ ID NO:2 from amino acid number 26 (Tyr) to amino acid number 235 (Ser). Within another embodiment, the isolated polypeptide disclosed above is as shown in SEQ ID NO:2 from amino acid number 26 (Tyr) to amino acid number 235 (Ser). Within another embodiment, the isolated polypeptide disclosed above encodes motifs 1, 2, 3, 4 and 5 spaced apart from N-terminus to C-terminus in a configuration M1-{25-26}-M2-{15}-M3-{11}-M4-{34-36}-M5.
Within another aspect, the present invention provides a method of producing a z219a polypeptide comprising: culturing a cell as disclosed above; and isolating the z219a polypeptide produced by the cell.
Within another aspect, the present invention provides a method of producing an antibody to z219a polypeptide comprising: inoculating an animal with a polypeptide selected from the group consisting of: (a) a polypeptide consisting of 9 to 210 amino acids, wherein the polypeptide is at least 90% identical to a contiguous sequence of amino acids in SEQ ID NO:2 from amino acid number 26 (Tyr) to amino acid number 235 (Ser); (b) a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 from amino acid number 26 (Tyr) to amino acid number 235 (Ser); (c) a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 from amino acid number 59 (Arg) to amino acid number 133 (Asp); (d) a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 from amino acid number 135 (Ser) to amino acid number 212 (Ala); (e) a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 from amino acid 215 (Asn) to amino acid number 231 (Pro); and wherein the polypeptide elicits an immune response in the animal to produce the antibody; and isolating the antibody from the animal.
Within another aspect, the present invention provides an antibody produced by the method disclosed above, which specifically binds to a z219a polypeptide. Within one embodiment, the antibody disclosed above is a monoclonal antibody.
Within another aspect, the present invention provides an antibody which specifically binds to a polypeptide disclosed above.
Within another aspect, the present invention provides a method of detecting, in a test sample, the presence of an antagonist of z219a protein activity, comprising: transfecting a z219a-responsive cell, with a reporter gene construct that is responsive to a z219a-stimulated cellular pathway; and producing a z219a polypeptide by the method disclosed above; and adding the z219a polypeptide to the cell, in the presence and absence of a test sample; and comparing levels of response to the z219a polypeptide, in the presence and absence of the test sample, by a biological or biochemical assay; and determining from the comparison, the presence of the antagonist of z219a activity in the test sample.
Within another aspect, the present invention provides a method of detecting, in a test sample, the presence of an agonist of z219a protein activity, comprising: transfecting a z219a-responsive cell, with a reporter gene construct that is responsive to a z219a-stimulated cellular pathway; and adding a test sample; and comparing levels of response in the presence and absence of the test sample, by a biological or biochemical assay; and determining from the comparison, the presence of the agonist of z219a activity in the test sample.
These and other aspects of the invention will become evident upon reference to the following detailed description of the invention and attached drawings.